HIV-1复制型DNA疫苗与非复制型重组痘苗病毒疫苗在小鼠体内细胞免疫效果的研究

摘要

Objective: In this research, we select HIV-1 replicating DNA vaccine and recombinant non-replicating Tiantan Vaccinia Virus (rNTV-C) vaccine to evaluate their immunogenicity in mice.The two vaccines respectively express six genes including gp 160, gag-pol and rev-tat-nef of HIV-1 clade B'/C.Methods: BALB/c mice were injected with HIV-1 DNA vaccine and rNTV-C alone or combined.The different injection dose and different injection pathway of the vaccine were designed.Cellular immune responses were detected with IFN-γ ELISPOT.Results: HIV-1 DNA vaccine and rNTV-C separated immunization could induce special cellular immune responses of antigens of HIV-1.The combined immunization of DNA priming and rNTV-C boost could elicit much higher cellular immune response than DNA or rNTV-C alone immunization.The statistical analysis results showed that the immune response of intramuscular(ira) injection of HIV-1 DNA vaccine and rNTV-C was higher than intradermal(id)injection.The immune response of 1 μg dose of DNA vaccine was similar with 5 μg dose and the immune response of l×l08 PFU dose of rNTV-C was higher than 2×l07 PFU dose.Conclusion: HIV-1 DNA vaccine and rNTV-C alone or combined immunization can induce special cellular immune response.The research data also provided important evidence for institution of clinical immune plan.%目的:用HIV-1复制型DNA疫苗和非复制型重组痘苗病毒疫苗(rNTV-C)进行单独免疫和联合免疫的研究(2种疫苗分别包含HIV-1 B'/C亚型的gp160、gag-pol、rev-tat-nef等6种基因),以了解这2种新型HIV疫苗单独免疫及联合免疫的效果,并为临床免疫方案的制定提供实验依据.方法:将HIV一1 DNA疫苗和rNTV-C疫苗免疫BALB/c小鼠,设计rNTV-C单独免疫组、DNA单独免疫组,以及DNA初免、rNTV-C加强的联合免疫组,并设计不同免疫途径和不同剂量的各种组合.用IFN-γ ELISPOT检测各组的细胞免疫效果,用统计学方法分析比较各组细胞免疫效果的差异.结果:DNA疫苗和rNTv-C疫苗单独免疫时,二者都能诱发针对各抗原的特异性免疫反应;联合免疫能够诱发比DNA或rNTV-C单独免疫都强的特异性细胞免疫反应.统计分析显示,2种疫苗采用肌肉注射途径的免疫效果显著高于皮内注射,1μg和5 μg DNA疫苗的免疫效果差异不显著,而lx108 PFU的rNTV-C比2x107PFU的免疫效果要强.结论:联合免疫策略能够显著增强HIV-1疫苗各抗原的免疫原性,通过对2种HIV-1疫苗单独免疫及二者联合免疫的细胞免疫反应的分析比较,确定了较好的免疫方案,为疫苗临床前免疫效果评价和临床方案的制定提供了实验依据.